Characterization of in Vitro Transdermal Iontophoretic Delivery of Insulin

In-vitro studies were conducted to characterize the transdermal iontophoretic delivery of insulin, thus avoiding potential complications from various biological variations, which may be encountered during in-vivo studies. The proteolytic degradation of insulin in skin homogenates and degradation under the experimental conditions used was investigated. Appropriate adjuvants were incorporated to minimize the potential degradation problems of insulin. ¹²⁵I-Insulin was observed to penetrate into and accumulate in the skin during the iontophoresis period. It was then released gradually from this depot, as a mixture of intact and ¹²⁵-I labelled fragments, into the receptor medium. Drug desorption studies supported the theory of skin depot or reservoir formation. It was found that an electric field could be used to facilitate the desorption of drug from the depot. The post-application flux of insulin (or its fragments) from the skin depot formed during iontophoresis was monitored to study the factors affecting the iontophoretic delivery of insulin. Stripping and delipidization of the skin were noted to increase the skin permeation rate of insulin. The cumulative radioactivity permeated and accumulated in the skin was higher at pH 3.6 than at pH 7.4. The iontophoresis-facilitated transdermal delivery was observed to increase with increasing duration of current application and increasing donor concentration of insulin. Modulation of drug delivery by multiple applications was also found to be feasible.     

Role of ATRX in chromatin structure and function: implications for chromosome instability and human disease

Functional differentiation of chromatin structure is essential for the control of gene expression, nuclear architecture, and chromosome stability. Compelling evidence indicates that alterations in chromatin remodeling proteins play an important role in the pathogenesis of human disease. Among these, α-thalassemia mental retardation X-linked protein (ATRX) has recently emerged as a critical factor involved in heterochromatin formation at mammalian centromeres and telomeres as well as facultative heterochromatin on the murine inactive X chromosome. Mutations in human ATRX result in an X-linked neurodevelopmental condition with various degrees of gonadal dysgenesis (ATRX syndrome). Patients with ATRX syndrome may exhibit skewed X chromosome inactivation (XCI) patterns, and ATRX-deficient mice exhibit abnormal imprinted XCI in the trophoblast cell line. Non-random or skewed XCI can potentially affect both the onset and severity of X-linked disease. Notably, failure to establish epigenetic modifications associated with the inactive X chromosome (Xi) results in several conditions that exhibit genomic and chromosome instability such as fragile X syndrome as well as cancer development. Insight into the molecular mechanisms of ATRX function and its interacting partners in different tissues will no doubt contribute to our understanding of the pathogenesis of ATRX syndrome as well as the epigenetic origins of aneuploidy. In turn, this knowledge will be essential for the identification of novel drug targets and diagnostic tools for cancer progression as well as the therapeutic management of global epigenetic changes commonly associated with malignant neoplastic transformation.     

Einfluß des Spannungsarmglühens auf die mechanischen Eigenschaften und das Gefüge von schweißsimulierend wärmebehandelten Proben aus warmfesten Feinkornbaustählen

Untersuchung der Eigenschaften des grobkörnigen Gefüges im wärmebeeinflußten Bereich neben einer Schweißnaht an simulierend wärmebehandelten Proben von sechs Feinkornbaustählen, nämlich 17 MnMoV 6 4, 17 MnNiMoV 5 4, 17 MnNiMoNb 6 6, 22 NiMoCr 3 7, 17 MnNiMo 5 4 und 17 NiCrMo 10 6. Einfluß des Spannungsarmglühens vorwiegend bei 580 °C bis 100 h Dauer auf Härte und Kerbschlagzähigkeit. Ermittlung der Übergangstemperatur für die Kerbschlagzähigkeit, der Bruchzähigkeit und der Rißfortschrittgeschwindigkeit. Zeitstandversuche bis 100 h Dauer vorwiegend bei 580 °C an glatten und gekerbten Proben sowie Entspannungsversuche an glatten Proben. Zusammenhang zwischen Zeitstandkerbempfindlichkeit, Zeitbruchdehnung, Entspannungsverhalten und Rißbildung bei Spannungsarmglühen. Abgrenzung der Bedingungen für das Entstehen von Rissen beim Spannungsarmglühen.     

Reproducibility of in vivo carotid intima-media thickness measurements: a review

The gyrA gene of Campylobacter fetus subsp. fetus, which encodes the A subunit of DNA gyrase, was cloned, and its nucleotide sequence was determined. An open reading frame of 2,586 nucleotides which encodes a polypeptide of 862 amino acids with an Mr of 96,782 was identified. C. fetus subsp. fetus GyrA is most closely related to Campylobacter jejuni GyrA, with 73% homology at the nucleotide level and 78% identity between polypeptides. The next most closely related GyrA was that from Helicobacter pylori, with both DNA homology and amino acid identity of 63%. The gyrA and gyrB (DNA gyrase B subunit) genes were located on the genomic map of C. fetus subsp. fetus ATCC 27374 and shown to be separate. A clinical isolate of C. fetus subsp. fetus and a laboratory-derived mutant of ATCC 27374, both resistant to ciprofloxacin, had identical mutations within the quinolone resistance determining region. In both mutants a G-->T transversion, corresponding to a substitution of Asp-91 to Tyr in GyrA, was linked to ciprofloxacin resistance, giving MICs of 8 to 16 micrograms/ml.     

TGFβ-Treated Placenta-Derived Mesenchymal Stem Cells Selectively Promote Anti-Adipogenesis in Thyroid-Associated Ophthalmopathy

Orbital fibroblasts (OFs) in thyroid-associated ophthalmopathy (TAO) are differentiated from pre-adipocytes and mature adipocytes; increased lipid and fat expansion are the major characteristics of ophthalmic manifestations. Human placental mesenchymal stem cells (hPMSCs) were reported to immunomodulate pathogenesis and suppress adipogenesis in TAO OFs. Here, we prepared transforming growth factor β (TGFβ, 20 ng/mL)-treated hPMSCs (TGFβ-hPMSCs) in order to enhance anti-adipogenic effects in vitro and in TAO mice. TAO OFs were grown in a differentiation medium and then co-cultured with hPMSCs or TGFβ-hPMSCs. TAO OFs were analyzed via quantitative real-time polymerase chain reaction, Oil red O staining, and western blotting. The results showed that TGFβ-hPMSCs reduced the expression of adipogenic, lipogenic, and fibrotic genes better than hPMSCs in TAO OFs. Moreover, the adipose area decreased more in TAO mice injected with TGFβ-hPMSCs compared to those injected with hPMSCs or a steroid. Further, TGFβ-hPMSCs inhibited inflammation as effectively as a steroid. In conclusion, TGFβ-hPMSCs suppressed adipogenesis and lipogenesis in vitro and in TAO mice, and the effects were mediated by the SMAD 2/3 pathways. Furthermore, TGFβ-hPMSCs exhibited anti-inflammatory and anti-fibrotic functions, which suggests that they could be a new and safe method to promote the anti-adipogenic function of hPMSCs to treat TAO patients.     

Human B cells secreting immunoglobulin G to glutamic acid decarboxylase-65 from a nondiabetic patient with multiple autoantibodies and Graves' disease: a comparison with those present in type 1 diabetes

Antibodies to glutamic acid decarboxylase-65 (GAD65) are present in a number of autoimmune disorders, such as insulin-dependent (type 1) diabetes mellitus (IDDM), stiff man syndrome, and polyendocrine autoimmune disease. Antibodies to GAD in IDDM patients usually recognize conformation-dependent regions on GAD65 and rarely bind to the second isoform, glutamic acid decarboxylase-67 (GAD67). In contrast, those present in stiff man syndrome and polyendocrine disease commonly target the second isoform (GAD67) and include antibodies that are less dependent on the conformation of the molecule. By immortalizing peripheral blood B cells with Epstein-Barr virus, we have generated three human IgG autoantibodies, termed b35, b78, and b96, to GAD65 from one patient with multiple autoantibodies to endocrine organs and Graves' disease. All three autoantibodies are of the IgG1 isotype, with islet cell activity, and do not react with GAD67. The regions on GAD65 recognized by the three autoantibodies have been investigated by immunoprecipitation with a series of chimeras, by binding to denatured and reduced antigens, and using protein footprinting techniques. Using chimeric GAD proteins, we have shown that b35 targets the IDDM-E1 region of GAD65 (amino acids 240-435) whereas both b78 and b96 target the IDDM-E2 region of GAD65 (amino acids 451-570). Furthermore, examination of binding to recombinant GAD65 and GAD67 by Western blotting revealed some differences in epitope recognition, where only b78 bound denatured and reduced GAD65. However, b35, b78, and b96 autoantibodies had different footprinting patterns after trypsin treatment of immune complexes with GAD65, again indicating different epitope recognition. Our results indicate that antibodies to GAD65 present in nondiabetic patients with multiple autoantibodies to endocrine organs show similarities to those in IDDM (by targeting IDDM-E1 and IDDM-E2 regions of GAD65) as well as subtle differences in epitope recognition (such as binding to denatured and reduced GAD65 and by protein footprinting). Thus, the GAD65 epitopes recognized by autoantibodies in different autoimmune diseases may overlap and be more heterogeneous than previously recognized.     

Active cellular preform derived from stems of jute and sticks of cane and their suitability for bulk porous Si/SiC ceramics

Stems of Jute (Corchorus capsularis L.) and sticks of Cane (Calamus rotang L.), plants of immense economic importance in the Indian subcontinent, were converted into carbonaceous perform (C-preform) maintaining the circular cylindrical shapes in lengths of 0.02–0.05 m by controlled thermal processing. Plant material precursors were characterized by analysis of elemental (C, H, N) and molecular (cellulose, hemicellulose, lignin) compositions, by determination of Bulk Density (BD) and ash content and by optical microscopy and X-ray diffractometry (XRD). C-preforms were also characterized by measurement of BD and by Scanning Electron Microscopy (SEM) and XRD. The C-preforms were further subjected to infiltration with Si-melt (1823–1923 K) under vacuum. Spontaneous infiltration and reaction yielded composite ceramics preserving the morphology of native Jute Stem (JS) and Cane Stick (CS) precursors on macro and micro scale. The materials were found to be duplex composites with Si and β-SiC as crystalline phases. The end ceramics were characterized by measurement of BD, and also by SEM and by XRD. Measured mean BD of the Si/SiC composites derived from JS and CS were 2190 Kg m⁻³ and 2250 Kg m⁻³. The respective volume fractions of large diameter (>100 μm) bulk pores were 0.134 and 0.204, in the composites derived from JS and CS. Taking into account the measured volume fraction internal pores of 0.11 and 0.149, the volume fractions of SiC were calculated to be 0.136 and 0.307 in the composites derived from JS and CS respectively, closely tallying with those calculated from the C-preform bulk densities. The cellular Si/SiC ceramics derived from JS and CS having special morphologies with long and large porous channels and oriented growth of constituent phases are likely to be suitable for devices such as high temperature insulators, catalyst support structures for gas phase reactions at elevated temperatures, molten metal filters and others.